TY - JOUR
T1 - Secreted factors from mouse embryonic fibroblasts maintain repopulating function of single cultured hematopoietic stem cells
AU - Marquez, Romero
AU - Hettler, Franziska
AU - Hausinger, Renate
AU - Schreck, Christina
AU - Landspersky, Theresa
AU - Henkel, Lynette
AU - Angerpointner, Corinne
AU - Demir, Ihsan E.
AU - Schiemann, Matthias
AU - Bassermann, Florian
AU - Götze, Katharina S.
AU - Istvánffy, Rouzanna
AU - Oostendorp, Robert A.J.
N1 - Publisher Copyright:
© 2021 Ferrata Storti Foundation
PY - 2021/10
Y1 - 2021/10
N2 - Hematopoietic stem cell self-renewal, proliferation, and differentiation are independently regulated by intrinsic as well as extrinsic mechanisms. We previously demonstrated that proliferation of murine hematopoietic stem cells is supported in serum-free medium supplemented with two growth factors, stem cell factor and interleukin 11. The survival of hematopoietic stem cells is additionally improved by supplementing this medium with two more growth factors, neural growth factor and collagen 1 (four growth factors) or serum-free medium conditioned by the hematopoietic stem cell-supportive stromal UG26-1B6 cells.1 Here, we describe a robust and versatile alternative source of conditioned medium from mouse embryonic fibroblasts. We found that this conditioned medium supports survival and phenotypic identity of hematopoietic stem cells, as well as cell cycle entry in single cell cultures of CD34- CD48- CD150+ Lineage- SCA1+ KIT+ cells supplemented with two growth factors. Strikingly, in comparison with cultures in serum-free medium with four growth factors, conditioned medium from mouse embryonic fibroblasts increased the numbers of proliferating clones and the number of Lineage- SCA1+ KIT+ cells, with both two and four growth factors. In addition, conditioned medium from mouse embryonic fibroblasts supported self-renewal in culture of cells with short- and long-term hematopoiesis-repopulating ability in vivo. These findings identify conditioned medium from mouse embryonic fibroblasts as a robust, alternative, serum-free source of factors to maintain self-renewal of in vivo-repopulating hematopoetic stem cells in culture.
AB - Hematopoietic stem cell self-renewal, proliferation, and differentiation are independently regulated by intrinsic as well as extrinsic mechanisms. We previously demonstrated that proliferation of murine hematopoietic stem cells is supported in serum-free medium supplemented with two growth factors, stem cell factor and interleukin 11. The survival of hematopoietic stem cells is additionally improved by supplementing this medium with two more growth factors, neural growth factor and collagen 1 (four growth factors) or serum-free medium conditioned by the hematopoietic stem cell-supportive stromal UG26-1B6 cells.1 Here, we describe a robust and versatile alternative source of conditioned medium from mouse embryonic fibroblasts. We found that this conditioned medium supports survival and phenotypic identity of hematopoietic stem cells, as well as cell cycle entry in single cell cultures of CD34- CD48- CD150+ Lineage- SCA1+ KIT+ cells supplemented with two growth factors. Strikingly, in comparison with cultures in serum-free medium with four growth factors, conditioned medium from mouse embryonic fibroblasts increased the numbers of proliferating clones and the number of Lineage- SCA1+ KIT+ cells, with both two and four growth factors. In addition, conditioned medium from mouse embryonic fibroblasts supported self-renewal in culture of cells with short- and long-term hematopoiesis-repopulating ability in vivo. These findings identify conditioned medium from mouse embryonic fibroblasts as a robust, alternative, serum-free source of factors to maintain self-renewal of in vivo-repopulating hematopoetic stem cells in culture.
UR - http://www.scopus.com/inward/record.url?scp=85116228682&partnerID=8YFLogxK
U2 - 10.3324/haematol.2020.249102
DO - 10.3324/haematol.2020.249102
M3 - Article
C2 - 33543864
AN - SCOPUS:85116228682
SN - 0390-6078
VL - 106
SP - 2633
EP - 2640
JO - Haematologica
JF - Haematologica
IS - 10
ER -